Molecular Biology Core

分子生物学，从广义上讲，涵盖了DNA和RNA的操作，已成为大多数生物学研究计划的一个组成部分。利用分子生物学技术的研究领域包括克隆基因以在细胞模型系统中表达蛋白质，定点诱变和标记用于细胞定位和功能研究，基因分型，RNA分离和表达分析，以及转基因小鼠的开发。分子生物学核心于1998年在北卡罗来纳州（北卡罗来纳州）囊性纤维化中心大学成立，为通常主要接受细胞生物学，生理学或临床科学培训的教师提供分子生物学服务和专业知识。核心项目目前是项目编制补助金中所述的许多正在进行的项目的一个组成部分，提供专家指导、技术支持和专门设备。PPG应用程序中的所有四个项目都将使用分子生物学核心服务。PPG研究人员将需要将基因克隆到逆转录病毒、慢病毒和卵母细胞表达载体中，以在各种细胞模型中表达各种标记和/或修饰的蛋白质（CFTR、ENaC亚基和核苷转运蛋白），包括分化良好的人支气管上皮细胞（HBE）（由Core C：Cell Culture提供）。其次，将提供RNA分离和表达分析服务，包括定量实时PCR和si/shRNA试剂的开发和测试，沿着更专业的服务，如RNA测序和数据分析。最后，分子生物学核心将有助于产生和利用新的转基因小鼠模型，即，Panxl和Slc 17 A9，以帮助识别气道表面液体稳态的重要调节因子。分子生物学核心将有效地补充项目负责人及其工作人员的专业知识，并使他们能够专注于自己个人精通领域的实验。

Molecular biology, which in a broad sense covers the manipulation of DNA and RNA, has become an integral part of most biological research programs. Research areas that utilize molecular biology techniques include cloning genes for expression of proteins in cell model systems, site-directed mutagenesis and tagging for cellular localization and functional studies, genotyping, RNA isolation and expression analysis, and the development of transgenic mice. The Molecular Biology Core was established at the University of North Carolina (UNC) Cystic Fibrosis Center in 1998 to provide molecular biology services and expertise to faculty who are often primarily trained in cell biology, physiology, or clinical sciences. The Core is currently an integral part of many of the ongoing projects described in the PPG providing expert guidance, technical support, and specialized equipment. All four projects within the PPG application will utilize Molecular Biology Core services. PPG investigators will require cloning of genes into retroviral, lentiviral and oocyte expression vectors for expression of a variety of tagged and/or modified proteins (CFTR, ENaC subunits, and nucleoside tranporters) in a variety of cell models, including well-differentiated human bronchial epithelial (HBE) (provided by Core C:Cell Culture). Secondly, RNA isolation and expression analysis services, including quantitative real-time PCR and development and testing of si/shRNA reagents, will be provided, along with more specialized services, such as RNA-sequencing and data analyses. Finally, the Molecular Biology Core will aid in the generation and utilization of new transgenic mouse models, i.e., Panxl and Slc17A9, to help identify important regulators of airway surface liquid homeostasis. The Molecular Biology Core will effectively complement the expertise of project leaders and their staff and to allow them to focus on the experiments in their own personal areas of proficiency.